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Biomedical subjects

K Tamura

Publications and source records attributed to K Tamura.

At least 433 records · Page 24Linked to original sources

Induction of additional limb at the dorsal-ventral boundary of a chick embryo.

In the early chick embryo, an apical ectodermal ridge (AER) is formed from the overlying ectoderm of the presumptive limb bud region at the dorsal-ventral (DV) boundary. We report here that the ectopic DV boundary formed in the presumptive wing, flank, and leg fields induces an ectopic AER structure. Dorsal tissue (ectoderm and mesoderm) from the presumptive wing field of stage 10 to 17 embryos was inserted into a slit in the somatopleure of the future ventral side of host embryos. The same method was used to implant ventral tissue into the future dorsal side of host embryos. After the implantation, ectopic AER was induced and an additional limb or limb-like structure developed. In related experiments, ectoderm-free presumptive wing tissue was implanted, which resulted in a considerably decreased frequency of ectopic AER formation. Further analysis of chick and quail chimeras suggests that the ectopic AER was formed from the ectodermal cells overlying the boundary of host and graft mesodermal cells. These results indicate that the DV boundary organizes the AER structure in the limb bud field of early-stage chick embryos and that the ectoderm of the grafted tissues plays an important role in this process.

Animals↗

Effects of hyperthermal stress on the ultrastructure of platelets with reference to the localization of platelet peroxidase and fibrinogen in vivo.

Ultrastructure of platelets with the localization of platelet peroxidase and fibrinogen through 3-min 47 degrees C hot-spring bathing was investigated in eight healthy volunteers. The mean sublingual temperature rose about 1.8 degrees C 5 min after the start of bathing. The frequencies of fold, pseudopods, vacuoles, and centralization were increased after bathing. Platelet peroxidase activity was decreased after bathing. Furthermore, fibrinogen was decreased in alpha-granules after bathing. Thus, hyperthermal stress in vivo may activate platelets, resulting in consumption of platelet peroxidase and fibrinogen.

Adult↗

Ascidian tyrosinase gene: its unique structure and expression in the developing brain.

Tadpole larvae of ascidians have two sensory pigment cells in the brain. One is the otolith cell that functions as a gravity receptor, the other pigment cell is part of a primitive photosensory structure termed the ocellus. These sensory cells, like vertebrate pigment cells, contain membrane-bounded melanin granules and are considered to reflect a crucial position in the evolutionary process of this cell type. To investigate the molecular changes accompanying the evolution of pigment cells, we have isolated from Halocynthia roretzi a gene encoding tyrosinase, a key enzyme in melanin biosynthesis. The cDNA has an open reading frame (ORF) of 596 amino acids, which is 36-39% identical in amino acid sequence to vertebrate tyrosinases. In addition, the sequence analysis of both cDNA and genomic clones reveals an unusual organization of the tyrosinase gene, an extraordinary 3' untranslated region of the transcripts with significant homology to the coding sequence, and a single short intron in the sequence encoding a cytoplasmic domain. Expression of the gene is detected first in two pigment precursor cells positioned in the neural plate of early neurulae, and later in two melanin-containing pigment cells within the brain of late tailbud embryos. Its expression pattern correlates well with the appearance of tyrosinase enzyme activity in the developing brain. These results provide the first description of pigment cell differentiation at the molecular level in the ascidian embryo, and also will contribute to a better understanding of the evolution of chordate pigment cells.

Amino Acid Sequence↗

Shh expression in developing and regenerating limb buds of Xenopus laevis.

The zone of polarizing activity (ZPA) is a specialized region involved in the antero-posterior (A-P) axis formation in chick and mouse limb buds. The existence of ZPA in the posterior margin is suggested in Xenopus hindlimb buds because 180 degree rotation of the distal limb tip induces the supernumerary limb. In this study, we investigated the expression of Sonic hedgehog (shh), a molecular marker for ZPA, in Xenopus developing limb buds and regenerating blastemas by whole-mount in situ hybridization. Although shh was expressed in the posterior margin of the limb bud like in chicks, its expression domain did not correspond to the ZPA map of Xenopus hindlimb buds. shh expression was distant from the ZPA at stage 53 in particular. To clarify the difference between the shh expression domain and ZPA, we examined shh expression in 180 degree rotated limb buds. As a result, ectopic shh expression was newly induced in the proximal region to its original expression domain. These results suggest that ZPA is accompanied by shh expression as in chick limb buds. Furthermore we examined shh expression in regenerating blastemas. shh was reexpressed in the posterior margin of the blastema. This result supports the possibility that ZPA also exists in the regenerating blastema.

Animals↗

Resolution of late potentials with improvement in left ventricular systolic function in patients with first acute myocardial infarction.

BACKGROUND: Ventricular late potentials predict subsequent arrhythmic events and sudden death in postinfarction patients. Late potentials are recorded in the infarcted area, but it should be pointed out that they probably represent micropotentials in the area of delayed conduction found among isolated areas of scar tissue and normal myocardium. HYPOTHESIS: The study was undertaken to investigate the relationship between chronic reversible myocardial ischemia, such as stunned or hibernating myocardium, and late potentials in 38 patients with a first myocardial infarction. METHODS: The patients were divided into two groups, one with (Group 1) and one without (Group 2) resolution of late potentials recorded by signal-averaged electrocardiogram at 6 months after onset of myocardial infarction. We investigated the clinical, echocardiographic, and signal-averaged electrocardiographic characteristics of Groups 1 and 2. RESULTS: In the chronic phase of myocardial infarction, a higher incidence of patency of the infarct-related artery and an improvement of wall motion score were found in Group 1, and left ventricular ejection fraction was better preserved in Group 1 than in Group 2. CONCLUSIONS: These results suggest that the resolution of late potentials was influenced by the improvement of left ventricular systolic function and patency of the infarct-related artery. We concluded that chronic reversible myocardial ischemia, such as stunned or hibernating myocardium, might be the substrate that generated late potentials.

Action Potentials↗

Experimental investigation of the role of the splenic vein in gastric venous obstruction after pylorus-preserving pancreatoduodenectomy in the dog.

CONCLUSION: This study showed that the splenic vein (SV) plays a major role in maintaining gastric blood flow after ligature of peripyloric vessels in the dog. Therefore, SV preservation or alternative venous drainage may be recommended to maintain sufficient gastric blood flow in pylorus-preserving pancreatoduodenectomy (PPPD). BACKGROUND: In PPPD, we often encounter the problem of gastric venous drainage obstruction, if the splenic-portal vein confluence needs to be resected. This study investigated significance of venous drainage via the left gastric and splenic veins in gastric tissue blood flow. METHODS: In seven mongrel dogs, gastric tissue blood flow was measured by both laser Doppler flowmetry (LDF) in the antrum and hydrogen gas clearance (HGC) in the antrum and body. Blood flow was measured after the following treatments: 1. No treatment; 2. Duodenal transection; 3. Ligature of the right gastric and gastroepiploic arteries and veins; 4. Transient obstruction of the left gastric vein (LGV); 5. Transient obstruction of the SV with the LGV open; and 6. Obstruction of both the LGV and the SV. RESULTS: As judged by LDF, treatments 2, 3, and 4 did not decrease blood flow, but treatments 5 and 6 significantly decreased it compared with both treatments 1 and 3. By HGC, changes in the antrum and the body were parallel. Duodenal transection did not, but treatments 3 and 4 did decrease blood flow compared with no treatment. Obstruction of the left gastric vein did not, but treatments 5 and 6 did significantly decrease it compared with treatment 3.

Animals↗

Mucosal blood flow and generation of superoxide in rat experimental colitis induced by succinic acid.

As we consider succinic acid to be an exacerbating factor in ulcerative colitis, we investigated its influence on rat colonic mucosa in terms of mucosal blood flow and superoxide generation. We measured mucosal blood flow by the hydrogen gas clearance method and superoxide generation by the chemiluminescence method, and observed histopathological findings to determine the effects of succinic acid. After the instillation of succinic acid of any concentration tested to the colon, mucosal blood flow decreased. Histopathologically, the higher the concentration of succinic acid, the greater was the erosion formation in the colonic mucosa, while significant polymorpho-nuclear cell infiltration superoxide generation from colon tissue were observed with 0.01% succinic acid compared with higher or lower concentrations. Succinic acid, at fecal concentrations found in active stage ulcerative colitis, appears to be implicated in mucosal injury, mediated by a decrease in colonic mucosal blood flow and infiltration of superoxide-generating polymorpho-nuclear cells into the mucosa.

Animals↗

The effects of transmural pressure on prostacyclin release from porcine endocardial endothelial cells--comparison with vascular endothelial cells.

We assessed the effect of pressure on the release of prostacyclin (PGI2) from cultured endocardial endothelial cells (EECs) and vascular endothelial cells (VECs). EECs were harvested from the right ventricle (RV) and left ventricle (LV) of porcine hearts, and VECs from pulmonary artery (PA), aorta (Ao) and coronary artery (CA). Confluent EECs and VECs were incubated for 30 min under various pressures (0, 50, 100, 150 mmHg) and PGI2 release from each cell was measured. Pressure-induced PGI2 release from LV-EECs was larger than that from RV-EECs. Pressure also increased PGI2 release from both PA- and Ao-VECs, but not from CA-VECs. These findings suggest that endocardium can produce PGI2 in response to pressure and PGI2 released into the coronary blood from the ventricle may play an important role in the prevention of myocardial ischemia.

Animals↗

Germline p53 mutation at codon 133 in a cancer-prone family.

We identified four families in which we suspected the presence of genetic factors predisposing them to cancer. We examined one family with features suggesting Li-Fraumeni syndrome for the presence of a germline p53 mutation in 13 of its members. To detect germline p53 mutations we performed polymerase chain reaction/nonradioisotopic single-strand conformation polymorphism and DNA sequencing analysis on exons 4-9 of the p53 gene. Mutated polymerase chain reaction-restriction fragment length polymorphism analysis was also performed on exon 5 to confirm the mutation identified by the sequencing analysis. A novel germline p53 mutation was identified at codon 133 (ATG-->AGG) in exon 5, resulting in the substitution of arginine for methionine, in all four cancer-affected individuals and in three apparently healthy individuals. We also analyzed tumor specimens for additional p53 mutations in the wild-type alleles using the same methods. However, heterozygosity was retained, and no other additional mutations in the wild-type allele were identified in any of the tumor tissues. It is possible that additional mutations in the wild-type allele are not always necessary for the loss of tumor suppressor functions. This study presents serious clinical and ethical problems about the predictive value of identifying germline p53 mutations in presymptomatic carriers. However, accurate predictive testing will be very useful in identifying unaffected individuals who are at increased risk of developing cancer and in detecting cancer at an early stage.

Adult↗

Amino acid losses and nitrogen balance during slow diurnal hemodialysis in critically ill patients with renal failure.

OBJECTIVE: The effects of slow diurnal hemodialysis (slow HD) on amino acid losses and nitrogen balance were studied. DESIGN: Slow HD was conducted for 10 h during the day at the dialysate flow rate of 30 ml/min. The patients received total parenteral nutrition including 40 g of amino acids (6.08 g of nitrogen). The amino acid concentrations in plasma and dialysate were determined and the daily nitrogen balance was calculated from the urea nitrogen appearance. PATIENTS: Six critically ill patients with renal failure were entered into the study. RESULTS: Slow HD eliminated 48.5 +/- 4.4 mmol (6.2 +/- 0.6 g) of amino acids, representing 16% of the daily amino acid load. The estimated nitrogen balance was -2.3 +/- 1.3 g/day. Amino acid nitrogen lost in the dialysate was 1.0 +/- 0.1 g, contributing 43% of the daily negative nitrogen balance. CONCLUSION: The amount of amino acid losses during slow HD should be taken into consideration when designing nutritional schedules for maintaining positive nitrogen balance in critically ill patients.

Aged↗

Antithrombotic effects of endocardial endothelial cells-comparison with coronary artery endothelial cells.

The purpose of this study was to assess the anti-platelet properties of endocardial endothelial cells (EECs) by measuring platelet aggregation after a brief incubation with cultured EECs. EECs were isolated from the right ventricles of porcine hearts and coronary artery endothelial cells (C-ECs) were also isolated from the same animals. After brief incubations (2-min) of platelet suspensions with cultured EEC and CEC monolayers, platelet aggregation in response to thrombin and 6-keto-PGF1 alpha (a stable metabolite of PGI2) content of platelet suspensions were measured. Platelet aggregation was significantly inhibited by a brief incubation of platelet suspensions with EEC and C-ECs monolayers. Pretreatment of EECs and C-ECs with indomethacin (5 x 10(-5) M) restored platelet activity, but pretreatment with N omega-nitro-L-arginine methyl ester (L-NAME, 5 x 10(-5) M) or hemoglobin (1 x 10(-6) M) did not. Platelet/EEC interactions multiplicatively increased the 6-keto-PGF1 alpha content of platelet suspensions and the 6-keto-PGF1 alpha content of platelet suspensions after incubations with EECs correlated significantly with the inhibition of platelet aggregation. Both the anti-aggregation properties and 6-keto-PGF1 alpha production were significantly greater in EECs than in C-ECs. A brief incubation (2-min) with PDGF (10 ng/ml) or TGF-beta (1 and 10 ng/ml) stimulated 6-keto-PGF1 alpha production in EECs but not in C-ECs, although these growth factors stimulated 6-keto-PGF1 alpha production in C-ECs after a longer incubation time (30 or 60 min). In this study, after a brief incubation (2-min) with platelet suspensions, EECs inhibited platelet aggregation mainly through the release of PGI2 but not EDRF. As this anti-aggregation property was significantly greater in EECs than in C-ECs, it is suggested that endocardial endothelial PGI2 may inhibit both intracardiac and intracoronary artery thrombus formation, contributing to the prevention of myocardial ischemia.

6-Ketoprostaglandin F1 alpha↗

Association of preceding angina with in-hospital life-threatening ventricular tachyarrhythmias and late potentials in patients with a first acute myocardial infarction.

We studied 140 patients with a first acute myocardial infarction to examine the effect of preceding angina as a marker of ischemic preconditioning on clinical ventricular arrhythmias and late potentials. Preceding angina was defined as the presence of ischemic chest pain within 24 hours before onset of myocardial infarction lasting no longer than 30 minutes and seen three or more times per day or at rest. Clinical features, angiographic findings, and late potentials were compared between patients with and without preceding angina. Thirty-four (24%) patients had preceding angina. Although the incidence of life-threatening ventricular tachyarrhythmias significantly differed (p = 0.0219), other clinical findings, including presence of late potentials, were not different between the two groups. Of 14 patients with life-threatening ventricular tachyarrhythmias, five events were considered as reperfusion arrhythmias. In patients who had successful reperfusion therapy, the incidence of life-threatening ventricular tachyarrhythmias had a tendency to be lower in patients with preceding angina than in those without preceding angina (p = 0.0586). Severe angina within 24 hours of onset of acute myocardial infarction is suggested to reduce occurrence of life-threatening ventricular tachyarrhythmias mainly associated with reperfusion during hospitalization.

Action Potentials↗

Hydroxyl radical scavenging effect of nicaraven in myocardial and coronary endothelial preservation and reperfusion injury.

OBJECTIVE: We investigated the efficacy of nicaraven in reducing myocardial as well as coronary endothelial preservation-reperfusion (P/R) injury. METHODS: In experiment I, isolated rat hearts were mounted on a Langendorff (L) apparatus to estimate the baseline cardiac function. Group 1, 8- and 12-h storage in histidine-tryptophan-ketoglutarate (HTK) solution; group 2, 8- and 12-h storage in HTK solution with superoxide dismutase (2.5 x 10(5) U/I) and catalase (2 x 10(5) U/I); group 3, 8- and 12-h storage in HTK solution with nicaraven (10(-3) M). Following storage for 8 and 12 h at 4 degrees C, they were reperfused and post-preservative cardiac function was evaluated. The hearts were then switched back to L-mode and paced at 330 beats/min. Coronary flow (CF) following perfusion with KHB solution containing 5-hydroxytryptamine (5-HT) and nitroglycerin was also measured. In experiment 2, segments of pig coronary artery were suspended in organ chambers and exposed to hydroxyl radicals in the presence or absence of nicaraven. The sensitivity of relaxation response to bradykinin of the prior-exposed rings was measured. RESULTS: The recovery of CF and LV dp/dt following 8 h of storage in group 3 was higher than that in group 1, although there were no significant differences in the other parameters of cardiac functional recovery among the groups. The absolute values of cardiac function following 12 h of storage in groups 1-3 were as follows: 6.6 +/- 2.4, 9.1 +/- 0.8, 15.6 +/- 3.1 ml/min of cardiac output (CO); 1.9 +/- 0.9, 2.3 +/- 0.4, 6.0 +/- 2.3 ml/min of aortic flow (AF); 4.5 +/- 1.3, 6.0 +/- 0.5, 9.5 +/- 0.8 ml/min of CF, respectively. The recovery of CO, AF, CF, SP, MP, and left ventricular (LV) dp/dt were significantly improved in group 3, compared with those in group 1. CF and CO in group 3 were higher than those in group 2.5-HT caused vasoconstriction in all groups, but the vasoconstriction in group 3 was less than in group 1. Prior exposure to FeSO4/H2O2 produced significant endothelial damage as reflected by the right-ward shift of the dose-response curve of bradykinin-induced endothelium-dependent relaxation. In the presence on nicaraven, the dose-response curve recovered to the control level. CONCLUSIONS: Nicaraven may improve coronary endothelial and myocardial function following P/R by its hydroxyl radical scavenging activity.

Animals↗

New indices of ischemic heart disease and aging: studies on the serum levels of soluble intercellular adhesion molecule-1 (ICAM-1) and soluble vascular cell adhesion molecule-1 (VCAM-1) in patients with hypercholesterolemia and ischemic heart disease.

It is known that the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on the surface of vascular endothelial cells is closely related to the formation of early atherosclerotic lesions. In this study, serum soluble ICAM-1(sICAM-1) and soluble VCAM-1(sVCAM-1) were determined by sandwich ELISA both in normal healthy individuals (n = 114) and in patients with hypercholesterolemia (HC, n = 112) or ischemic heart disease (IHD, n = 38) to clarify the significance of the soluble forms of the adhesion molecules in the development of atherosclerotic diseases. IHD patients, not HC patients, showed significant elevation of sICAM-1, but not of sVCAM-1, compared with controls in age and sex-matched subjects. In addition, multiple linear regression analysis showed that sICAM-1 was correlated only to the presence of IHD but not to age and lipids. Multiple logistic regression analysis revealed that sICAM-1 was the most powerful independent predictor of the presence of IHD. On the other hand, sVCAM-1, not sICAM-1, was positively correlated to age. Multiple linear regression analysis showed that age was the most powerful independent predictor of the level of sVCAM-1. These data suggest that sICAM-1 and sVCAM-1 are useful as indices of clinical manifestations of atherosclerosis and aging, respectively.

Adult↗

Relationship between hepatic angiotensinogen mRNA expression and plasma angiotensinogen in patients with chronic hepatitis.

Recent association and linkage studies suggested that angiotensinogen may play an important role in the pathogenasis of essential hypertension. However, there is little information in human concerning a relationship between plasma angiotensinogen levels and the angiotensinogen mRNA expression in the liver, which is the main production site of angiotensinogen. Therefore, the aim of this study was to examine whether hepatic angiotensinogen gene expression determines the level of circulating angiotensinogen and the activity of the renin-angiotensin system in humans. The subjects were 36 patients with chronic hepatitis. Blood was collected from each patients for estimation of plasma renin activity, plasma angiotensinogen and angiotensin II concentrations and several parameters of liver function. In addition, total RNA was isolated from liver biopsy specimens, which were then used to measure angiotensinogen mRNA with Northern blot analysis. Levels of angiotensinogen mRNA were detected easily in the liver biopsy specimens in all of the patients. Hepatic angiotensinogen mRNA levels were positively correlated with plasma angiotensinogen levels (r=0.41, P=0.013). In contrast, hepatic angiotensinogen mRNA levels did not show any significant relationship with plasma renin activity, plasma angiotensin II concentration, histological subgroup of hepatitis, histological activity index and parameters of liver function tests. The present study demonstrated, for the first time, that hepatic angiotensinogen mRNA levels correlated with plasma angiotensinogen concentration in humans.

Adult↗

Dogs develop resistance to Rhipicephalus sanguineus.

The capacity of Beagle dogs to develop resistance against infestation by adult females of Rhipicephalus sanguineus was investigated. The number of successive engorged females recovered from dogs at the second infestation of the tick was significantly less than at the first infestation. However, there were no significant differences in body weight, body length, body width of engorged females, feeding behavior, preoviposition periods or egg weight.

Animals↗

Direct observation of the separation process by the chromato-videoscope.

Separation process in a liquid chromatographic column were visualized and analyzed by a developed chromato-videoscope. The migration aspects were evaluated with successively obtained densitograms. In reversed-phase chromatography, the band widths of each solute band were almost equal fore both weakly and strongly retained solutes when compared at the same column position (the same migration distance in the column). In the gradient elution mode, the position of the solute band showed that the migration velocity of the solute band changed gradually according to changes in solvent composition. The drug trapping process to BSA- coated ODS packings for direct injection of biological fluid was also observed. In the absence of BSA from the sample solution, the drug molecules were trapped in a narrow band. However, at higher BSA concentrations in the sample solution, a broader band shape was observed. This band broadening shows how the drug molecules were retained on the protein.

Chromatography, High Pressure Liquid↗